ALTERNATIVE TRANSCIPTION OF HUMAN ALTERNATIVE TRANSCIPTION OF HUMAN CHM GENE INCLUDING LTR12C ELEMENT IN COLORECTAL CHM GENE INCLUDING LTR12C ELEMENT IN COLORECTAL

CANCERCANCER Yi-Deun Jung1, Yun-Ji Kim1, Jae-Won Huh2, Dae-Soo Kim3, Hong-Seok Ha1, Eun-Sang Choi1,

Won-Ho Lee1 and Heui-Soo Kim1

1 Division of Biological Sciences, College of Natural Sciences, Pusan National University, Busan 609-735, Republic of Korea2 National Primate Research Center, KRIBB, Ochang, Chungbuk 363-883, Republic of Korea

3 Korea Bioinformation Center, KRIBB, Daejeon 305-806, Republic of Korea

purewonder@pusan.ac.kr Abstract

CHM (Choroideremia) is indispensable for the posttranslational activation of retina-specific Rab protein. Various mutations such as missense, nonsense, rearrangement of the CHM gene are responsible for choroideremia. We could find several elements including MIR, LINE, Alu and LTR12C that are located within the intron 4 of the CHM gene. Alternatively spliced transcript including LTR12C element was detected in the last exon of gene. In order to detect this transcript, we accompanied RT-PCR analysis in the normal and adjacent cancer tissue samples, indicating that this transcript was expressed higher in colorectal cancer than normal tissue samples with similar expression patterns in other cancer tissues (liver, lung, testes). We also confirmed high expression of colorectal cancer in patient sample. The expression level of alternative transcript of CHM gene could be used to predict colorectal cancer. Further study about effect of LTR12C in this alternative transcript between normal and cancer tissues is indicated.

1.Patience C, Takeuchi Y, Weiss RA (1997) Infection of human cells by an endogenous retrovirus of pigs. Nat Med 3: 276-282. 2.Scheef G, Fischer N, Krach U, Tonjes RR (2001) The number of a U3 repeat box acting as an enhancer in long terminal repeats of polytropic replication-competent porcine endogenous retroviruses dynamically fluctuates during serial virus passages in human cells. J Virol 75: 6933-6940

Introduction Materials & Methods

Results & Discussion

References

TSD TSDLTR gag pol env LTRPBS

gag : coding for matrix and capsid proteinspol : reverse transcriptase and integrase proteins

env : product viral envelope and transmembrane structures

LTR : long terminal repeatLINE : long integrated nuclear elementSINE : short integrated nuclear elementHERV : Human endogenous retro-virus

8%

RNase

Viral RNA

Capsid : core proteins

Receptorbinding proteins

Lipid envelope

• Human & Primate genomic DNA

Bioinformatics ( in silico analysis CHM gene )

Genomic DNA PCR & Gene Cloning

RT PCR ( Human original & EST form)

colorectal tissue samples RT-PCR, Image J analysis

CHM ( choroideremia )Chr X

q21.21 2 3 4 5 6 7 8 9 1011 12 13 14 15

85120551 85120427 85105713 85105326

4 5

CDSUTR MIR/SINE Alu/SINE L1/LINE ERV1/LTR trigger5

1 2 3 4 5

Alternative splicing transcriptEST

Original

RT-PCR (original transcript & EST form)1 2 3 4 5 6 7 8 9 10M

11 12 13 14 15 16 17 18 19 20M

1 : adrenal gland, 2 : bone marrow, 3 : adult brain cerebellum, 4 : adult brain whole, 5 : fetal brain, 6 : fetal liver, 7 : heart, 8 : kidney, 9 : liver, 10 : lung, 11 : placenta, 12 :

salivary gland, 13 : SK-muscle, 14 : spinal cord, 15 : testis, 16 : prostate, 17 : thymus, 18 : thyroid, 19 : trachea, 20 : uterus

N C N C N

Transcript including LTR12C RT-PCR (colorectal cancer tissue) EST

Original

C N C N C N C

N C N C N C N C

Analyzed quantitatively using the Image-J program